Abstract

Abstract Executioner caspases, such as caspase‐3, are known to induce apoptosis, but in other contexts, they can control very different fates, including cell differentiation and neuronal plasticity. While hundreds of caspase substrates are known to be specifically targeted during cell death, we know very little about how caspase activity brings about non‐apoptotic fates. Here, we report the first proteome identification of cleavage events in C2C12 cells undergoing myogenic differentiation and its comparison to undifferentiated or dying C2C12 cells. These data have identified new caspase substrates, including caspase substrates specifically associated with differentiation, and show that caspases are regulating proteins involved in myogenesis in myotubes, several days after caspase‐3 initiated differentiation. Cytoskeletal proteins emerged as a major group of non‐apoptotic caspase substrates. We also identified proteins with well‐established roles in muscle differentiation as substrates cleaved in differentiating cells.